title: Feeder layer- and animal product-free culture of neonatal foreskin keratinocytes: improved performance, usability, quality and safety.
abstract: Since 1987, keratinocytes have been cultured at the Queen Astrid Military Hospital. These keratinocytes have been used routinely as auto and allografts on more than 1,000 patients, primarily to accelerate the healing of burns and chronic wounds. Initially the method of Rheinwald and Green was used to prepare cultured epithelial autografts, starting from skin samples from burn patients and using animal-derived feeder layers and media containing animal-derived products. More recently we systematically optimised our production system to accommodate scientific advances and legal changes. An important step was the removal of the mouse fibroblast feeder layer from the cell culture system. Thereafter we introduced neonatal foreskin keratinocytes (NFK) as source of cultured epithelial allografts, which significantly increased the consistency and the reliability of our cell production. NFK master and working cell banks were established, which were extensively screened and characterised. An ISO 9001 certified Quality Management System (QMS) governs all aspects of testing, validation and traceability. Finally, as far as possible, animal components were systematically removed from the cell culture environment. Today, quality controlled allograft production batches are routine and, due to efficient cryopreservation, stocks are created for off-the-shelf use. These optimisations have significantly increased the performance, usability, quality and safety of our allografts. This paper describes, in detail, our current cryopreserved allograft production process.
authors.affiliation: Skin- and Keratinocyte Bank, Laboratory for Molecular and Cellular Technology, Burn Wound Centre, Queen Astrid Military Hospital, Bruynstraat 1, 1120 Brussels, Belgium.
authors.forename: Peter
authors.initials: P
authors.lastname: De Corte
authors.forename: Gunther
authors.initials: G
authors.lastname: Verween
authors.forename: Gilbert
authors.initials: G
authors.lastname: Verbeken
authors.forename: Thomas
authors.initials: T
authors.lastname: Rose
authors.forename: Serge
authors.initials: S
authors.lastname: Jennes
authors.forename: Arlette
authors.initials: A
authors.lastname: De Coninck
authors.forename: Diane
authors.initials: D
authors.lastname: Roseeuw
authors.forename: Alain
authors.initials: A
authors.lastname: Vanderkelen
authors.forename: Eric
authors.initials: E
authors.lastname: Kets
authors.forename: David
authors.initials: D
authors.lastname: Haddow
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Pirnay
mesh_terms: D000818:Animals; D001706:Biopsy; D018929:Cell Culture Techniques / Q000379:methods* / Q000592:standards*; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D061252:Feeder Cells / Q000166:cytology*; D052816:Foreskin / Q000166:cytology* / Q000637:transplantation; D006801:Humans; D007231:Infant, Newborn; D015603:Keratinocytes / Q000166:cytology* / Q000637:transplantation; D008297:Male; D051379:Mice; D012449:Safety; D014015:Tissue Banks; D014019:Tissue Donors; D014184:Transplantation, Homologous